LOCUS       pEF.DEST51-PTPN1        8943 bp DNA     circular SYN 26-AUG-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8943)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 20, 2024 from SnapGene Viewer 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8943
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        475..1653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          706..1644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1670..1688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1720..1744
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1745..1763
                     /label=MCS 1
                     /note="multiple cloning site, part 1"
     CDS             1807..4137
                     /codon_start=1
                     /label=PTPN12(D199A,C231S)
                     /label=PTPN12
                     /translation="MEQVEILRKFIQRVQAMKSPDHNGEDNFARDFMRLRRLSTKYRTE
                     KIYPTATGEKEENVKKNRYKDILPFDHSRVKLTLKTPSQDSDYINANFIKGVYGPKAYV
                     ATQGPLANTVIDFWRMIWEYNVVIIVMACREFEMGRKKCERYWPLYGEDPITFAPFKIS
                     CEDEQARTDYFIRTLLLEFQNESRRLYQFHYVNWPAHDVPSSFDSILDMISLMRKYQEH
                     EDVPICIHSSAGCGRTGAICAIDYTWNLLKAGKIPEEFNVFNLIQEMRTQRHSAVQTKE
                     QYELVHRAIAQLFEKQLQLYEIHGAQKIADGVNEINTENMVSSIEPEKQDSPPPKPPRT
                     RSCLVEGDAKEEILQPPEPHPVPPILTPSPPSAFPTVTTVWQDNDRYHPKPVLHMVSSE
                     QHSADLNRNYSKSTELPGKNESTIEQIDKKLERNLSFEIKKVPLQEGPKSFDGNTLLNR
                     GHAIKIKSASPCIADKISKPQELSSDLNVGDTSQNSCVDCSVTQSNKVSVTPPEESQNS
                     DTPPRPDRLPLDEKGHVTWSFHGPENAIPIPDLSEGNSSDINYQTRKTVSLTPSPTTQV
                     ETPDLVDHDNTSPLFRTPLSFTNPLHSDDSDSDERNSDGAVTQNKTNISTASATVSAAT
                     STESISTRKVLPMSIARHNIAGTTHSGAEKDVDVSEDSPPPLPERTPESFVLASEHNTP
                     VRSEWSELQSQERSEQKKSEGLITSENEKCDHPAGGIHYEMCIECPPTFSDKREQISEN
                     PTEATDIGFGNRCGKPKGPRDPPS"
     misc_feature    2401..2403
                     /label=double-mutant, D199A
     misc_feature    2497..2499
                     /label=Catalytic Cysteine, C231S
     CDS             join(4147..4149,4150..4152,4153..4863)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     misc_feature    4873..4875
                     /label=MCS 2
                     /note="multiple cloning site, part 2"
     protein_bind    complement(4879..4903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4929..4970
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4980..4997
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5026..5250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5296..5724
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5738..6068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5919..6054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6116..6163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6182..6580
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6738..6859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(7310..7898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(8069..8860,8861..8929))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(8930..8943,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgcctc gtgaggctcc
      481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     1021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     1081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1621 caaagttttt ttcttccatt tcaggtgtcg tgaggaatta gcttggtact aatacgactc
     1681 actataggga gacccaagct ggctaggtaa gcttgatcaa caagtttgta caaaaaagca
     1741 ggctggcgcc ggaaccaatt cagtcgactg gatccggtac cgaggagatc tgccgccgcg
     1801 atcgccatgg agcaagtgga gatcctgagg aaattcatcc agagggtcca ggccatgaag
     1861 agtcctgacc acaatgggga ggacaacttc gcccgggact tcatgcggtt aagaagattg
     1921 tctaccaaat atagaacaga aaagatatat cccacagcca ctggagaaaa agaagaaaat
     1981 gttaaaaaga acagatacaa ggacatactg ccatttgatc acagccgagt taaattgaca
     2041 ttaaagactc cttcacaaga ttcagactat atcaatgcaa attttataaa gggcgtctat
     2101 gggccaaaag catatgtagc aactcaagga cctttagcaa atacagtaat agatttttgg
     2161 aggatgatat gggagtataa tgttgtgatc attgtaatgg cctgccgaga atttgagatg
     2221 ggaaggaaaa aatgtgagcg ctattggcct ttgtatggag aagaccccat aacgtttgca
     2281 ccatttaaaa tttcttgtga ggatgaacaa gcaagaacag actacttcat caggacactc
     2341 ttacttgaat ttcaaaatga atctcgtagg ctgtatcagt ttcattatgt gaactggcca
     2401 gcccatgatg ttccttcatc atttgattct attctggaca tgataagctt aatgaggaaa
     2461 tatcaagaac atgaagatgt tcctatttgt attcatagca gtgcaggctg tggaagaaca
     2521 ggtgccattt gtgccataga ttatacgtgg aatttactaa aagctgggaa aataccagag
     2581 gaatttaatg tatttaattt aatacaagaa atgagaacac aaaggcattc tgcagtacaa
     2641 acaaaggagc aatatgaact tgttcataga gctattgccc aactgtttga aaaacagcta
     2701 caactatatg aaattcatgg agctcagaaa attgctgatg gagtgaatga aattaacact
     2761 gaaaacatgg tcagctccat agagcctgaa aaacaagatt ctcctcctcc aaaaccacca
     2821 aggacccgca gttgccttgt tgaaggggat gctaaagaag aaatactgca gccaccggaa
     2881 cctcatccag tgccacccat cttgacacct tctccccctt cagcttttcc aacagtcact
     2941 actgtgtggc aggacaatga tagataccat ccaaagccag tgttgcatat ggtttcatca
     3001 gaacaacatt cagcagacct caacagaaac tatagtaaat caacagaact tccagggaaa
     3061 aatgaatcaa caattgaaca gatagataaa aaattggaac gaaatttaag ttttgagatt
     3121 aagaaggtcc ctctccaaga gggaccaaaa agttttgatg ggaacacact tttgaatagg
     3181 ggacatgcaa ttaaaattaa atctgcttca ccttgtatag ctgataaaat ctctaagcca
     3241 caggaattaa gttcagatct aaatgtcggt gatacttccc agaattcttg tgtggactgc
     3301 agtgtaacac aatcaaacaa agtttcagtt actccaccag aagaatccca gaattcagac
     3361 acacctccaa ggccagaccg cttgcctctt gatgagaaag gacatgtaac gtggtcattt
     3421 catggacctg aaaatgccat acccatacct gatttatctg aaggcaattc ctcagatatc
     3481 aactatcaaa ctaggaaaac tgtgagttta acaccaagtc ctacaacaca agttgaaaca
     3541 cctgatcttg tggatcatga taacacttca ccactcttca gaacacccct cagttttact
     3601 aatccacttc actctgatga ctcagactca gatgaaagaa actctgatgg tgctgtgacc
     3661 cagaataaaa ctaatatttc aacagcaagt gccacagttt ctgctgccac tagtactgaa
     3721 agcatttcta ctaggaaagt attgccaatg tccattgcta gacataatat agcaggaaca
     3781 acacattcag gtgctgaaaa agatgttgat gttagtgaag attcacctcc tcccctacct
     3841 gaaagaactc ctgaatcgtt tgtgttagca agtgaacata atacacctgt aagatcggaa
     3901 tggagtgaac ttcaaagtca ggaacgatct gaacaaaaaa agtctgaagg cttgataacc
     3961 tctgaaaatg agaaatgtga tcatccagcg ggaggtattc actatgaaat gtgcatagaa
     4021 tgtccaccta ctttcagtga caagagagaa caaatatcag aaaatccaac agaagccaca
     4081 gatattggtt ttggtaatcg atgtggaaaa cccaaaggac caagagatcc accttcatgc
     4141 ggccgcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4201 ctggacggcg acgtaaacgg ccacaagttc agcgtccgcg gcgagggcga gggcgatgcc
     4261 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     4321 cccaccctcg tgaccacctt cggctacggc gtggcctgct tcagccgcta ccccgaccac
     4381 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4441 atctctttca aggacgacgg tacctacaag acccgcgccg aggtgaagtt cgagggcgac
     4501 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4561 gggcacaagc tggagtacaa cttcaacagc cacaacgtct atatcacggc cgacaagcag
     4621 aagaacggca tcaaggctaa cttcaagatc cgccacaacg ttgaggacgg cagcgtgcag
     4681 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     4741 aaccactacc tgagccatca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     4801 atggtcctgc tggagttcgt gaccgccgcc gggattacac atggcatgga cgagctgtac
     4861 aagtagtaag atatctagac ccagctttct tgtacaaagt ggttgatcta gagggcccgc
     4921 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc
     4981 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     5041 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5101 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5161 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5221 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5281 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     5461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5581 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5641 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     5701 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     5761 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5821 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5881 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5941 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6001 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6061 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     6121 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     6181 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     6241 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     6301 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     6361 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     6421 aaatgagaac aggggcatct tgagcccctg cggacggtgt cgacaggtgc ttctcgatct
     6481 gcatcctggg atcaaagcga tagtgaagga cagtgatgga cagccgacgg cagttgggat
     6541 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc
     6601 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6661 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6721 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6781 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6841 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6901 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6961 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7501 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7741 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8041 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8101 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8161 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8221 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8281 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8341 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8401 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8461 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8521 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8581 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8641 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8701 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8761 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8821 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8881 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8941 ttc
//
